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Two and three‐dimensional gene transfer from enzymatically degradable hydrogel scaffolds
Author(s) -
Lei Yuguo,
Ng Quinn K.T.,
Segura Tatiana
Publication year - 2010
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20840
Subject(s) - self healing hydrogels , luciferase , chemistry , ethylene glycol , mesenchymal stem cell , tissue engineering , microbiology and biotechnology , gene transfer , gene delivery , biophysics , dna , in vivo , gene expression , green fluorescent protein , gene , transfection , biochemistry , biology , polymer chemistry , genetics , organic chemistry
The ability to genetically modify mesenchymal stem cells (MSCs) seeded inside synthetic hydrogel scaffolds would offer an alternative approach to guide MSC differentiation. In this report, we explored gene transfer to MSCs seeded on top or inside matrix metalloproteinase (MMP) degradable hydrogels that were loaded with DNA/poly(ethylene imine) (PEI) polyplexes. DNA/PEI polyplexes were encapsulated inside poly(ethylene glycol) (PEG) hydrogels crosslinked with MMP degradable peptides via Michael Addition chemistry. Gene transfer was visualized and quantified through using a vector encoding for green fluorescent protein and luciferase. We found that gene transfer to MSCs was possible for cells seeded both in two and three dimensions. The amount of luciferase expression was similar for cells seeded in two and three dimensions even though the number of cells in three dimensions is significantly higher, indicating that gene transfer to cells seeded in two dimensions is more efficient than for cells seeded in three dimensions. The use of hydrogel scaffolds that allow cellular infiltration to deliver DNA may result in long‐lasting signals in vivo, which are essential for the regeneration of functional tissues. Microsc. Res. Tech. 73:910–917, 2010. © 2010 Wiley‐Liss, Inc.